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Genetic Dissection of Quantitative Trait in Yeast and Discovery of Single Feature Polymorphisms in Gene Expression Profiling Microarray Data

Acknowledgements第4-10页
LIST OF FIGURES第10-12页
LIST OF TABLES第12-13页
ABSTRACT第13-14页
摘要第15-16页
PART Ⅰ第16-115页
    CHAPTER 1.INTRODUCTION第17-45页
        1.1.Genetic Basis of Phenotype Variation第17-20页
        1.2.Identifying Genes Affecting Quantitative Traits第20-32页
            1.2.1.Mutagenesis第20-23页
            1.2.2.QTL Mapping第23-29页
                1.2.2.1.Basis Concepts of QTL mapping第23-26页
                1.2.2.2.Molecular Markers第26-27页
                1.2.2.3.High Resolution Mapping第27-28页
                1.2.2.4.From QTL to Gene第28-29页
            1.2.3.Genetical Genomics第29-32页
        1.3.Ethanol Tolerance in the Budding Yeast Saccharomyces cerevisiae第32-43页
            1.3.1.Yeast as a model organism第32-34页
            1.3.2.Ethanol tolerance in yeast第34-43页
                1.3.2.1.Ethanol Fermentation with Yeast第34-35页
                1.3.2.2.Biochemical/Physiological Determinants of Ethanol Tolerance in Yeast第35-40页
                1.3.2.3.Progress in Genetic Dissection of Ethanol Tolerance in Yeast第40-43页
        1.4.Objective第43-45页
    CHAPTER 2.SELECTION OF HIGHLY PHENOTYPICALLY DIVERGENT STRAINS第45-68页
        2.1.Reagents and Solutions第46-49页
            2.1.1.Reagents第46页
            2.1.2.Solutions第46-49页
        2.2.Materials第49-50页
            2.2.1.Strains第49页
            2.2.2.Plasmids第49-50页
        2.3.Methods第50-61页
            2.3.1.Phenotype Scoring of Ethanol Tolerance第50页
            2.3.2.Preparation of Competent Cells第50-51页
            2.3.3.Bacteria Transformation第51-52页
            2.3.4.Plasmid Extraction第52-53页
            2.3.5.Yeast Transformation第53-54页
            2.3.6.Yeast DNA Extraction第54-55页
            2.3.7.Yeast Tetrad Spores Dissection第55-56页
            2.3.8.HO Gene Knockout第56-57页
            2.3.9.Diploidization and Mating-type Switch第57-58页
            2.3.10.Selection of Parent Strains with Divergent Ethanol Tolerance Phenotype第58-61页
            2.3.11.Gene Dosage Assay第61页
        2.4.Results第61-66页
            2.4.1.Phenotype Distribution in 53 Yeast Strains第61-62页
            2.4.2.Selection for Parent Strain with Low Ethanol Tolerance第62-65页
            2.4.3.Performance of Haploid and Diploid Strains第65-66页
        2.5.Discussion第66-68页
    CHAPTER 3.GENOMEWIDE SCANNING FOR ET QTL AND CANDIDATE QTL GENES IN F_2 POPULATION第68-87页
        3.1.Reagents and Materials第69-70页
            3.1.1.Reagents第69-70页
            3.1.2.Materials第70页
        3.2.Methods第70-76页
            3.2.1.Establishment of F_2 Mapping Population第70-71页
            3.2.2.Screening of Short Tandem Repeat Markers第71页
            3.2.3.Screening of SNP Markers第71页
            3.2.4.Genotyping第71-72页
            3.2.5.DNA Gel Extraction第72-73页
            3.2.6.PCR Purification第73-74页
            3.2.7.DNA Sequencing第74-75页
            3.2.8.Mapping Ethanol-tolerance QTL第75-76页
        3.3.Results第76-85页
            3.3.1.F_2 Mapping Population第76-77页
            3.3.2.Molecular Marker第77-78页
            3.3.3.Genomewide Scanning for ET QTL and Candidate QTL Genes第78-81页
            3.3.4.Multi-locus Association Analysis第81-85页
        3.4.Discussion第85-87页
    CHAPTER 4.FINE MAPPING USING RECURRENT SELECTION AND BACKCROSS BREEDING SCHEME第87-108页
        4.1.Background第87-88页
        4.2.Theoretical Basis of RSB第88-90页
        4.3.Materials第90-91页
        4.4.Methods第91-94页
            4.4.1.Construction of Parent Strains with Opposite Mating Types第91页
            4.4.2.RSB Breeding Scheme第91-92页
            4.4.3.Genotyping of RSB Individuals第92-93页
            4.4.4.Test for Maker-ET Association in RSB Mapping Population第93-94页
        4.5.Results第94-104页
            4.5.1.Construction of RSB Populations第94-98页
                4.5.1.1.Selection of Highly Tolerant Segregants第94-95页
                4.5.1.2.Selection of Sensitive Segregants第95-96页
                4.5.1.3.Introgression of Donor Genes at Two STR Loci第96-98页
            4.5.2.Marker-ET Association第98-102页
            4.5.3.Fine Mapping on Chromosome 9 and Functional Evaluation of the Candidate Gene第102-104页
        4.6.Discussion第104-108页
    BIBLIOGRAPHY第108-115页
PART Ⅱ第115-172页
    CHAPTER 5.INTRODUCTION第116-129页
        5.1.Backgronud第116-119页
        5.2.DNA Microarray第119-123页
        5.3.Allelic Variation Scanning Using DNA Microarray第123页
        5.4.Simultaneous Genotyping and Gene Expression Measurement第123-126页
        5.5.Objective第126-129页
    CHAPTER 6.YEAST AND BARLEY MICROARRAY DATA第129-144页
        6.1.Materials and Datasets第129-130页
            6.1.1.Yeast Strains and RSB Mapping Population第129页
            6.1.2.Yeast Genomic DNA Hybridizaiton and RNA Profiling Microarray Data第129-130页
            6.1.3.Barley Expression Profiling Microarray Data第130页
            6.1.4.Sources of Check Data第130页
        6.2.Experiment Protocols for Yeast Genome 2.0 Arrays第130-144页
            6.2.1.Reagents第131-132页
            6.2.2.Labeled Genomic DNA Preparation第132-134页
                6.2.2.1.Preparation of Yeast Cells for DNA Extraction第132页
                6.2.2.2.Isolation of DNA from Yeast第132-133页
                6.2.2.3.Determination of Yield and Purity of the DNA第133页
                6.2.2.4.Genomic DNA Fragmentation第133-134页
                6.2.2.5.Terminal Labeling and Checking of Labeling Efficiency第134页
                6.2.2.6.Quantifying Labeled DNA第134页
            6.2.3.RNA Target Preparation第134-142页
                6.2.3.1.Preparation of Yeast Cells for RNA Extraction第134-135页
                6.2.3.2.Hot Phenol Extraction of RNA from Yeast第135-136页
                6.2.3.3.Purification of Total RNA from Yeast第136-137页
                6.2.3.4.Preparation of Poly-A RNA Controls for One-Cycle cDNA Synthesis第137页
                6.2.3.5.First-Strand cDNA Synthesis第137-138页
                6.2.3.6.Second-Strand cDNA Synthesis第138-139页
                6.2.3.7.Cleanup of Double-Stranded cDNA第139页
                6.2.3.8.Synthesis of Biotin-Labeled cRNA第139-140页
                6.2.3.9.Cleanup of Biotin-Labeled cRNA第140-141页
                6.2.3.10.Quantification of Biotin-Labeled cRNA第141页
                6.2.3.11.Fragmenting the cRNA for Target Preparation第141-142页
            6.2.4.Microarray Hybridization第142页
            6.2.5.Washing,Staining,and Scanning第142-144页
    CHAPTER 7.STATISTICAL INFERENCE OF SINGLE FEATURE POLYMORPHISMS第144-163页
        7.1 Analytical Model第144-147页
        7.2.SFP Genotyping第147-148页
        7.3.Calculation of True Discovery Rate and Rate of False Negative第148-150页
        7.4.Results第150-163页
            7.4.1.Predictability of PM Hybridization Intensity第151-152页
            7.4.2.Consistency in SFPs Predicted from Parallel DNA and RNA Datasets第152-154页
            7.4.3.Proportion of Differentially Expressed Genes第154-155页
            7.4.4.Mutual Predictability among Different Methods第155-157页
            7.4.5.Efficiency to Predict Sequence Polymorphisms第157-160页
            7.4.6.Genotyping and Genetic Map Construction Using SFP第160-163页
    CHAPTER 8.DISCUSSION第163-168页
    BIBLIOGRAPHY第168-172页
LIST OF PUBLICATIONS第172-173页

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